The liver is the only organ which produces proteins and antibodies, both of which play a very important roles in the defense of people against diseases, viruses, bacteria infections. This discovery reveals any protein that contains a healthy cell will properly can be used against any diseases, viruses, bacteria, deficiencies, inhibitors, prion in our body and thus from now on mankind will be protected Against all kinds of diseases, cancers, epidemics, viruses, bacteria and possibly blind, deaf, mute people may benefit.
The cells of the blood can be divided into: white blood cells, red blood cells and platelets.
Red blood cells, or erythrocytes are the most common type of blood cell and our principal means of delivering oxygen to the body tissues through the circulatory system (arteries). Red blood cells take up oxygen in the lungs and release it while circulating through the body's capillary vessels (the smallest structures that conduct blood), where they take up carbon dioxide, which is a waste product of metabolism, and take it to the lungs, to be discarded through the respiration. These cells are rich in hemoglobin, which is a molecule that contains iron and that can bind oxygen (and is responsible for the blood's red color).
The red blood cells develop in the bone marrow and circulate for about 100-120 days in the body before their components are recycled by macrophages.
Red blood cells do not participate in the immune system.
Platelets are cell fragments (that is, cells that do not have a nucleus, 2-3 μm in diameter, which are derived from fragmentation of precursor cells known as “megakaryocytes”. The average lifespan of a platelet is normally just 5 to 9 days. Platelets play a fundamental role in hemostasis with the formation of clots, but they do not participate in the immune system.
White blood cells, or leucocytes, are cells of the immune system involved in defending the body against both infectious disease and foreign materials. There are five different and diverse types of leukocytes exist, but they are all produced and derived from a multi potent cell in the bone marrow, known as a hematopoietic stem cell. Leukocytes are found throughout the body, not only in the blood and the lymphatic system.
The number of white blood cells in the blood is often an indicator of disease. There are normally between 5,000 to 10,000 white blood cells per mL. An increase in the number of leukocytes over the upper limits s called leukocytosis, and a decrease below the lower limit is called leukopenia.
Type of cell%Main targetsLifetimeNeutrophil54-62Bacteria and fungi6 hs to afew daysLymphocyte25-33B Lymphocytes (releases antibodies and assistWeeks to“activation” of T lymphocytes)yearsT Lymphocytes:Helper (activate and regulate T and Blymphocytes)Cytotoxic T lymphocytes CD8+ (virus-infectedand tumor cells)Gamma-delta T lymphocytes (suppressor Tlymphocytes) Returns the functioning of theimmune system back to normal operation afterinfection and prevents autoimmunityNatural killer T lymphocytes (virus-infected andtumor cells)Eosinophil1-6Larger parasites8-12 daysModulate allergic inflammatory responsesBasophil<1Release mediators (histamine) in inflammatoryHours toresponsedaysMonocyte 2-10Monocytes migrate from the bloodstream toHours toother tissues and differentiate into tissuedaysresident macrophages or dendritic cellsMacrophageNo*Phagocytosis (engulfment and digestion) ofactivated:cellular debris and pathogens, and stimulation ofdayslymphocytes and other immune cells thatimmature:respond to the pathogen.months toyears
Another important support role in transporting protein is Chylomicrons (Mainly Triglycerides which is considered very BAD according to a lot of publications and it may lead to Heart attack and Stroke. However, without triglycerides FATS and cholesterol cannot Move within the water-based solution of the Blood stream.
Chylomicrons are large lipoprotein particles that consist of triglycerides (85-92%), phospholipids (6-12%), cholesterol (1-3%) and proteins (1-2%) [1]. They transport dietary lipids from the intestines to other locations in the body. Chylomicrons are one of the five major groups of lipoproteins (chylomicrons, VLDL, IDL, LDL, HDL) that enable fats and cholesterol to move within the water-based solution of the bloodstream.
Function: Chylomicrons transport exogenous lipids to liver, adipose, cardiac, and skeletal muscle tissue, where their triglyceride components are unloaded by the activity of lipoprotein lipase. As a consequence, chylomicron remnants are left over and are taken up by the liver.
Origin
Chylomicrons are a type of lipoprotein produced in absorptive cells of small intestines, specifically, the epithelial cells within the villi of the duodenum.
Stages
There are three stages in the chylomicron's “life cycle”:
Nascent chylomicron
Mature chylomicron
Chylomicron remnant
Nascent Chylomicrons
Chylomicrons are created by the absorptive cells of the small intestine, known as enterocytes. They are relatively large, having a diameter of 75 to 1,200 nm. These nascent chylomicrons are released by exocytosis from enterocytes into lacteals, lymphatic vessels originating in the villi of the small intestine, and are then secreted into the bloodstream at the thoracic duct's connection with the left subclavian vein.
Nascent chylomicrons are primarily composed of triglycerides (85%) and contain some cholesterol and cholesteryl esters. The main apolipoprotein component is apolipoprotein B-48 (APOB48).
Mature Chylomicron
While circulating in lymph and blood, chylomicrons exchange components with high-density lipoproteins (HDL). The HDL donates apolipoprotein C-II (APOC2) and lipoprotein E (APOE) to the nascent chylomicron and thus converts it to a mature chylomicron (often referred to simply as “chylomicron”). APOC2 is the cofactor for lipoprotein lipase (LPL) activity.
Chylomicron Remnant
Once triglyceride stores are distributed, the chylomicron returns APOC2 to the HDL (but keeps APOE), and, thus, becomes a chylomicron remnant, now only 30-50 nm. APOB48 and APOE are important to identify the chylomicron remnant in the liver for endocytosis and breakdown.
References1.^ M Mahmood Hussain: “Review Article: A proposed model for the assembly of chylomicrons”; Arterosclerosis; Vol. 148; 2000; pages 1-15;
INVENTIONS: Several Manufacturing processes of a protein that contain one of the Healthy GOOD CELLS as described. Few of them are described here and the rest as we can separate Protein by Protein by different process, it can be combined from one protein with others to contain one of these Health Good Cells.
The present invention relates to a method of introduction of a healthy good human cells to eat up bad damaged or cells, comprising administering an effective amount of a healthy good protein containing transferin, alpha 1-antitrypsin, apolipoprotein A and human albumin.
By the present invention, damage to healthy human cells can be reduced by administering an effective amount of a healthy good protein containing ApoA1/2/4 and/or transferrin and/or alpha 1 antitrypsin and/or C1 esterase inhibitors and other inhibitors.
The present invention also involves a method of introduction of healthy goodies human cells to eat up bad damaged cells to reduce damage to healthy human cells, comprising administering an effective amount of a healthy good protein containing Factor II, Factor VII, Factor IX and Factor X in Prothrombin Complex Concentrate (ProthoRAAS®). The damage to healthy human cells can be reduced by administering an effective amount of a protein containing any one or more of: human albumin (AlbuRAAS®), immunoglobulin (GammaRAAS®), fibrinogen (FibroRAAS®), Factor VIII (HemoRAAS®), high concentrate fibrinogen (FibrinGluRAAS®), thrombin (ThrombiRAAS®), Hepatitis B and Immune Globulin (HBIG) (HepaRAAS®).
The present invention further involves a method of reducing damage to healthy human cells, comprising administering an effective amount of a healthy good protein containing one or more of: Anti thrombin III (AT-III), protein C, fibronectin, protein S, and protein M. An effective amount of two or more of the proteins described herein for use with the present invention can be used. The proteins include all currently known proteins and proteins yet to be discovered in Fraction III of plasma.
The method of the present invention further involves reducing damage to healthy human cells by administering an effective amount of any of: a macrophage, a neutrophil, a basophil, a lymphocyte and an eosinophil in white blood cells, red blood cells, platelets, chylomicrons, electrolyses, and peptides in humans or in animals or in chemicals or in substances from any source of materials obtaining by clone expressing to obtain the cells for further purification by rDNA, monoclonal, transgenic, or by any other methodology.
The more healthy good proteins that are used in effective amounts in a combination of proteins means that the combination will be more potent and effective than a single healthy good protein in the treatment of diseases and viruses or bacterial infections. The method of reducing damage to healthy human cells according to the present invention comprises any good healthy protein of, for example, AFOD RAAS 1-85 and AFCC RAAS 1-85 can be combined with any currently available and future developed drugs to enhance the efficacy, while reducing toxicity and the side effects caused by chemical drugs.
The method according to the present invention also involves introducing healthy goodies human cells to eat up bad damaged cells, the method comprising administering an effective amount of a protein containing at least one of the following apolipoproteins: ApoA1, ApoA2, ApoA4, Apo-B48, ApoB100, ApoCI, ApoCII, ApoCIII, Apo-D, Apo-E, Apo-H, and Apo(a), all of which contain good healthy cells. The protein further contains at least one of the following: alpha 1 antitrypsin (A1AT), transferrin, and human albumin, all of which contain good healthy cells.
The 3rd generation of APO purification.
The main difference is that urea is no longer required but need 2 steps of chromatography, as is shown in FIG. 1.    1. A method to purify APO from plasma fraction IV,            1) Fraction IV is resuspended in a buffer with pH 3.00-10.00, and the celite and other impurities were separated by press filter or centrifugation, the resulted supernatant was then collected,        2) The APO in the supernatant was then precipitated by adding NaCl and then was spin to collect the paste,        3) The resulted APO was then resuspended and filtered,        4) The resulted suspension was then underwent DEAE ion exchanging chromatography and butyl chromatography,            2. the fraction IV was resuspended in NaAc buffer with pH 3.00-10.00    3. the APO was precipitated by NaCl, pH 3.0-10.0, cool down to −1 to 1 C    4. the paste of APO can be resuspended in WFI or NaCl solution with pH 3.00-10.00 and 0-10 C    5. The resulted suspension is filtered with 0.45 um filter.    6. The chromatography in step 4 is Canion (DEAE) and butyl    7. The purification of APO by chromatography compromising,            Canion chromatography, adjust pH of filtered APO suspension to 3.0-10.0 and ionic strength to 15-25 mM, load on DEAE chromatography, low salt wash the DEAE chromatography, and then high salt elute the DEAE chromatography, collect the resulted APO elute,        Butyl chromatography, the resulted APO elute from DEAE chromatography is adjusted to pH 3.0-10.0 and low salt wash for impurities, WFI or alkaline buffer wash to collect APO enriched elute            8. The low salt buffer is a buffer containing Tris with pH 3.00-10.0, the high salt buffer is a buffer containing NaCl, the low salt elute buffer is a buffer containing Tris, the alkaline buffer is a buffer containing NaOH with pH 3.0-10.0    9. The resulted high purity of APO is then dialyzed and concentrated with virus inactivation, adding stabilizer and lyophilized.Process Nr 4.
A process that is separated the Fraction IV into 4 Proteins, as can be appreciated from FIG. 2:
1. Transferrin
2. Human Albumin
3. APO
4. Alpha 1 Antitrypsin (A1AT)
5. Transferrin+Human Albumin+APO+Immunoglobulin
6. Human Albumin+APO+Alpha 1 AntiStrepsin (A1AT)+Immunoglobulin
7. Human Albumin+APO+Immunoglobulin
8. Human Albumin+APO+Immunoglobulin+Transferrin+Antitrypsin
Or all products from Nr 5 to Nr 8 can be processed separately and put together at the Non Sterile Final Bulk-Sterile Filtration-Filling-Final Products.
1. Re suspension of fraction IV and pretreatment
1) Fraction IV is resuspended in a buffer with pH 3.00-10.00,
2) The celite and other impurities were separated by press filter of centrifugation; the resulted suspension was then collected,
3) The suspension was then treated with SD virus inactivation,
4) The resulted suspension was then subject to a canion chromatography like DEAE,
5) Proteins were eluted in different fractions,
6) The different eluted fractions were then further purified,
2. The fraction IV was dissolved in low temperature buffer to achieve a in homogenous suspension,
3. The celite in resulted suspension can be removed by press filter or centrifugation,
4. the suspension was then cleared by depth filter
5. the resulted suspension was then treated with Tween-80 and TNBP for virus inactivation at 25 C for 6 hours,
6. The resulted suspension was adjusted pH and ionic strength and then subjected to a canion chromatography like DEAE. The targeted proteins were then binding to the canion chromatography resin, which are transferrin, human albumin, APO and A1AT. The 1st elution was salt solution to elude the transferrin. The 2nd elution was then eluded by a high concentration salt solution, which was APO. The 3rd eluted fraction was human albumin by a low pH solution. Finally the 4th elution was A1AT which was eluted by a high concentration salt solution.7. The resulted various elution was then subjected to different chromatography for further purification to achieve a high purity. The 1st elution fraction was subjected to a CM chromatography. The 2nd elution fraction was subjected to a butyl chromatography. The 3rd elution fraction was subjected to a blue chromatography. The 4th elution fraction was subjected to a blue chromatography and a subsequent butyl chromatography.8. The resulted protein fractions were then dialyzed and concentrated. The pH was adjusted and stabilizer was then added.9. The resulted protein solutions were subjected to DV20 filtration for virus removal except human albumin.10. The human albumin could be virus inactivated by Double pasteurization.11. The resulted transferrin, APO, human albumin and A1AT can be filled.
AFCCRAAS 1: These processes of protein containing Healthy Good cells in Process 1 and Process 3 below are specially designed for Hemophilia A, B and WvB who have Been infected by HBV, HCV and specially HIV during the early of 1980 when effective process of inactivation of Enveloped viruses has not been introduced.
1. Process to separate Factor II, VII, IX, and X Transferrin, Human Albumin, APO and A1AT (ProthoRAAS®) from fraction III which also contain at least 20 additional proteins.
2. Process to separate Factor II, VII, IX, and X (ProthoRAAS®)+Human Albumin (AlbuRAAS®)+Immunoglobulin (GammaRAAS)
3. Process to separate Factor II, VII, IX and X from Cryopaste
4. Process to separate Factor II, VII, IX and X from Cryopaste+ATA1 APO+ Human Albumin (AlbuRAAS®) and Immunoglobulin (GammaRAAS)
5. Process to separate Thrombin (ThrombiRAAS®) from Fraction III
6. Process to combine all protein from Fraction III.
Description
                1. As can be seen in FIG. 3, this process describes a process to purify prothrombin complex from cryoprecipitaiton, which comprises,        1) Re-constitute cryopaste in buffer containing 3,000 U/kg heparin,        2) Adjust pH and temperature        3) Complete mix at room temperature for 3 to 5 hours        4) Filter the resulted suspension with 10CP+90SP filter        5) Solvent detergent virus inactivation of resulted suspension        6) weak anion exchange chromatography of SD virus inactivated suspension        7) Twice washing of weak anion exchange chromatography        8) Elute weak anion exchange chromatography 2 to 3 times        9) Collect the result elution and ultra-filter with 10K membrane        10) Adjust pH of resulted elution        11) Adjust the activity of human FIX in the resulted elution        12) Aseptic filtration and nano filtration for virus removal        13) Filling and lyophilizationDescription        2. As can be seen in FIG. 4, this process describes a process to purify prothrombin complex from fraction III, which comprises,        14) Re-constitute cryopaste in buffer,        15) Adjust pH and temperature        16) PEG precipitation of resulted fraction III suspension        17) Centrifugation and collect the supernant        18) Filter the resulted suspension with 10CP+90SP filter        19) Solvent detergent virus inactivation of resulted suspension        20) weak anion exchange chromatography of SD virus inactivated suspension        21) Twice washing of weak anion exchange chromatography        22) Elute weak anion exchange chromatography 2 to 3 times        23) Collect the result elution and ultra-filter with 10K membrane        24) Adjust pH of resulted elution        25) Adjust the activity of human FIX in the resulted elution        26) Aseptic filtration and nano filtration for virus removal        27) Filling and lyophilizationAFOD is High Density Lipoprotein (ApoA1):        
Reference is made to FIGS. 5 and 6. The three dots in our analysis of AFOD are all ApoA1. The difference showed in 2D electropherosis of FIG. 6 might be due to different isoform of ApoA1 or Apo in the Apo family.                1. HDL (ApoA1): AFOD RAAS 1 (Trade mark) contains purified ApoA1 in the process described Nr 1 (China Patent granted 200610147503.7) Nr1&N2 (US 2009/0286960 A1) Nr 3,4 (U.S. 61/457,380) with a purity of 96% and the products must be free of all HIV1,2, HCV, HBV viruses and contain very good level of High Density Lipoprotein (HDL) which is GOOD CHOLESTEROL, No value or very low value of VLDL (Very low density Lipoprotein) and no value or very low value of LDL (Low Density Lipoprotein), both of which are BAD CHOLESTEROL. Potential applications: Cholesterol, Angina, hyperlipidemia, Clean plaque, fat on liver, Life span, Control of Obesity, Hypertension, Prevention of Heart attack, Prevention of Stroke, Prevention of Paralysis due to the stroke and other potential indications as described.        2. AFODRAAS 2 (Human Albumin+ApoA1) In addition to current clinical applications for AlbuRAAS® Potential applications for trauma management/Arthritis/Schizophrenia/Depression/Certain types of cancers Lung, Pancreas, Kidney, Liver, Prostate, Breast and other potential indications        3. AFODRAAS 3 (Intravenous Immuno Globulin+ApoA1) for current clinical applications for GammaRAAS® and for potential applications for all blood (liquid) cancers as described in abstract)        4. AFODRAAS 4 (Factor VIII+ApoA1) for the current clinical applications of HemoRAAS® and for potential applications for Hepatitis B, Hepatitis C and HIV 1,2, Bleeding complications/Bone Surgery in Patients with Hemophilia A (orthopedics, liver, pancreas, cancer of the gastrointestinal tube and eventually build up Coagulation so patients will no longer need to use Factor VIII for hemophilia A and all solid tumor and blood cancers/        5. AFODRAAS5 (Prothombin Complex Concentrate+ApoA1) for the current clinical applications of ProthoRAAS® and for potential applications for Hepatitis B, Cirrhosis, and other hepatic trouble like biliary tree obstruction Hepatitis C, HIV 1,2 Bleeding/Bone Complications and surgeries for Hemophilia B and Hemophilia A with inhibitor and eventually build up coagulation so patients will no longer need to use Factor IX or PCC and all solid tumor and blood cancers        6. AFODRAAS6 (Thrombin+ApoA1) for the current clinical applications of ThrombiRAAS® and for potential applications for Gastric and duodenal ulcer, ulcers and other problems of colon (large intestine)        7. AFODRAAS7 (Fibrinogen+ApoA1) for the current clinical applications of FibroRAAS® and for potential applications for Trauma Management and other potential indications as described        8. AFODRAAS8 (Fibrin Sealant+ApoA1) for the current clinical applications of FibrinGluRAAS® and potential TOPICAL APPLICATIONS for ALL SOLID TUMOR CANCERS which can be operated and cancers have not been spread to other parts of body.        9. AFODRAAS 9 (ApoA1+Human Albumin (AlbuRAAS)+Alpha 1 Antitrypsin (A1AT)+Transferrin for all potential indications as described        10. AFODRAAS 10 (ApoA1+Human Albumin+Alpha 1 Antitrypsin (A1AT) for all potential indications as described.        11. AFODRAAS 11 (ApoA1+Human Albumin+Transferrin) for all potential indications as described.        12. AFODRAAS 12 (ApoA1+Alpha 1 Antitrypsin ((A1AT)) for all potential indications as described.        13. AFODRAAS 13 (ApoA1+Transferrin for all potential indications as described.        14. AFODRAAS 14 (Alpha 1 Antitrypsin ((A1AT))+Transferrin) for all potential indications as described.        15. AFODRAAS 15 (Transferrin) for All potential indications as described.        
In our analysis of Fraction IV suspension by 2 D electropherosis, as shown in FIG. 7, the following proteins were found in the Fraction IV suspension:
The main proteins found in Fraction IV suspension are:                Transferrin, Human Albumin, Alpha 1 AntiStrepsin, and ApoA1        
The rest of the proteins are: SEMENOGELIN-1, HAPTOGLOBIN, VIMENTIN, NESPRIN-2, INTERFERON ALPHA 1/13, HP PROTEIN, VITAMIN D-BINDING, ALPHA-FETOPROTEIN, CASK, AMYLOID PRECURSOR, NEUREXINS AND SYNDECANS.
SEMENOGELIN-1: The protein encoded by this gene is the predominant protein in semen
HAPTOGLOBIN: In Blood Plasma, Haptoglobin binds free hemoglobin (Hb) released from erythrocytes with high affinity and thereby inhibits its oxidative activity.
VIMENTIN is a member of the intermediate filament family of proteins that is especially found in connective tissue. They, along with microtubules and actin microfilaments, make up the cytoskeleton
THE NESPRINS are a family of proteins that are found primarily in the outer nuclear membrane. Nesprin-1 and Nesprin-2 bind to actin filaments.
VITAMIN-D BINDING PROTEIN belongs to the albumin gene family, together with Human serum albumin and alpha-fetoprotein. It is a multifunctional protein found in plasma, ascetic fluid, cerebrospinal fluid and on the surface of many cell types. It binds to Vitamin D and its plasma metabolites and transport them to target tissues.CASK PROTEIN Peripheral plasma membrane protein CASK is a protein that in humans is encoded by the CASK gene. This protein is a multi domain scaffolding protein with a role in synaptic transmembrane protein anchoring and ion channel trafficking. It interacts with the transcription factor TBR1 and binds to several cell-surface proteins including amyloid precursor protein, neurexins, and syndecans.
FIG. 8 relates to Q8 Gene Symbol=GC Vitamin D binding protein Precursor—Cask isoform 3 of Peripheral plasma membrane protein CASk—VIM Vimentin
FIG. 9 relates to Q13 Gene Symbol=CASK Isoform 3 of Peripheral plasma membrane protein CASK—HP HP protein
FIG. 10 relates to Q15 Gene Symbol—CASK Isoform 3 of Peripheral plasma membrane protein CASK—IFNA13; IFNA1 Interferon alpha-1/13
AFCC is Prothombin Complex Concentrate (ProthoRAAS®) a combination of blood clotting factors II, VII, IX and X or Factor IX.
Factor IX is one of several factors made in the liver with similar structural properties.
Together with factors II (Prothombin), VII (proconvertin) and X (Stuart-Prower factor), factor IX (antihaemophilic factor B) comprises the so called prothombin complex group of factors, also known as PPSB factors.
Owing to their similarity, the proteins in this group are usually isolated together in fraction III in the Cohn alcohol fractionation process.
They are then purified to give preparations containing all four factors, Prothombin complex concentrates (PCC).
ProthoRAAS are indicated currently for:                Haemophilia B—Prophylaxis, Bleeding, Surgery        Haemophilia A—Inhibitor treatment        Liver disease—Acute and chronic—active hepatitis, cirrhosis        Vitamin K deficiency—Oral anticoagulants, Obstructive jaundice, Malabsorption, changes in intestinal flora (antibiotics, Morbus Crohn, ulcerative colitis)        DIC (Disseminated Intravascular Coagulation)—Infection, Liver disease, obstetrical emergencies, surgical complications.        16. AFCC RAAS1 (Trade mark) (ProthoRAAS®+ApoA1+AT-III) for the current indications as above together with all potential indications as described.        17. AFCC RAAS 2 (Trade mark) (Prothombin Complex Concentrate (ProthoRAAS®)+ApoA1+Human Albumin (AlbuRAAS®)), for the current indications as above together with all potential indications as described.        18. AFCC RAAS 3 (Trade mark) (Prothombin Complex Concentrate (ProthoRAAS®)+ApoA1+Intravenous Immuno Globulin (GammaRAAS®)), for the current indications as above together with all potential indications as described.        19. AFCC RAAS 4 (Trade mark) (Prothombin Complex Concentrate (ProthoRAAS®)+ApoA1+Intravenous Immuno Globulin (GammaRAAS®), +Human Albumin (AlbuRAAS®)), for the current indications as above together with all potential indications as described.        20. AFCC RAAS 5 (Trade mark) (Prothombin Complex Concentrate (ProthoRAAS®)+ApoA1+Intravenous Immuno Globulin (GammaRAAS®), +Human Albumin (AlbuRAAS®) and Fibrinogen (FibroRAAS®)), for the current indications as above together with all potential indications as described.        21. AFCC RAAS 6 (Trade mark) (Prothombin Complex Concentrate (ProthoRAAS®)+ApoA1+Fibrinogen (FibroRAAS®)), for the current indications as above together with all potential indications as described.2D Electropherosis of PCC:        
Results of 2D eletropherosis of PCC not only the above four factors proteins but also show a lot more proteins (Dots) that are being identified, as can be seen in FIG. 11.
2D Electropherosis of Fr. III:
2D electropherosis of Fraction III like Fraction IV contains a lot more of proteins other than Thrombin, Prothombin Complex, as can be seen in FIG. 12. All these proteins are also being identified.
2D Electropherosis of Cryopaste:
2 D electropherosis of Cryopaste results also show some other proteins which are being identified Beside Fibrinogen and Factor VIII, as can be seen in FIG. 13.
In Vitro/Vivo Studies
1st Production of 10 Batches totaling 200 grams for use in vivo study of Rabbits at Fudan University, Shanghai, China.
The 1st 200 g AFOD was purified in East China University of Science and Technology using Process No. 1, a flow chart for which is presented in FIG. 14. The pilot production capability of that lab is about 5 kg per time. The purification process was modified according to their equipment on site but the flow was same as we have done in this December The major differences are as following                1) The input of production was 5 kg of Fraction IV paste and yield was about 25 g AFOD each time. Dr. Li Chun Zhou produced 10 lots there to get 200 g AFOD;        2) The column used for AFOD purification was about 5 L DEAE chromatography;        3) The centrifugation for collecting AFOD enriched pellet was swing basket rotor centrifuge        4) Standing precipitation was used to get rid of celite;        5) For filling, manual filling was adopted;        6) No virus inactivation was used in the whole process.        
The resulted AFOD was about 200 g for total 10 lots and all of AFOD were lyophilized.
The stabilizer used was mannitol and this product has been used to perform clinical studies on 52 rabbits in summer of 2008.
2nd Pilot Production of 3 Lots at Shanghai RAAS Blood Products Co Ltd 2000 vials of AFOD RAAS (about 200 g) have been used in the following in vitro studies:                1. Studies of 5 cancer cells line at RuiJin Hospital Shanghai China.        2. Studies of 3 cancer cells line at R/D Lab Shanghai RAAS.        3. Studies of Bacteria at Microbiology Lab, Shanghai RAAS        4. Studies of Viruses HIV1,2 at NAT Lab, Shanghai RAAS        
In the most recent pilot production, which was conducted in last December at our Plant, we totally conducted 3 lots of production. It was 25 kg Fraction IV for the 1st lot, 50 kg for the rest of 2 lots. The total yield was about 2000 vials of AFOD (about 200 g). There were about 440 bottles of lyophilized, 660 vials of liquid with human albumin for stabilizer and another 880 bottles of liquid with mannitol for stabilizer.                1) We use continuous tube centrifugation to collect the AFOD enriched pellet;        2) Using Filtration to get rid of celite;        3) A 15 L DEAD chromatography for AFOD purification;        4) Virus inactivation including SD and DV 20;        5) Automatic filling.        
AFCC RAAS 1: A current product of Shanghai RAAS Blood products Co Ltd approved for Sales in China and has been exported to a certain country around the globe. Product has been manufactured at large industrial scale.
Product has been used in the VITRO studies at Shanghai RAAS Blood Products Co Ltd                1. 3 Cancer Cell lines at R/D Lab        2. Bacteria at Microbiology Lab        3. HIV 1+2 Testing at NAT LabIn Vitro Studies of Cancer Cell Lines:        
Procedures to test Cancer cells at RuiJin Hospital in Shanghai, China.